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目的观察牛蒡子苷元(ARG)对人食管癌细胞(EC-1)增殖、凋亡的影响,并探讨其机制。方法将不同浓度的ARG作用于EC-1细胞,采用噻唑蓝(MTT)实验检测细胞增殖活性(ARG浓度分别为2.5、5.0、10.0、20.0、40.0mg/L,作用时间为24、48、72、96、120h),流式细胞术检测细胞凋亡率,琼脂糖凝胶电泳和DNA缺口末端标记法(TUNEL)检测细胞凋亡指数,免疫细胞化学法检测凋亡相关基因Bcl-2、Bax表达水平的改变(该3种检测均采用:ARG浓度10.0、20.0、40.0mg/L,作用时间24h);分析不同浓度ARG对EC-1细胞的影响。并设阴性对照组(只加培养液不加药物)进行比较。结果 ARG对EC-1细胞增殖、凋亡的影响呈剂量/时间依赖性。随ARG浓度和作用时间的递增,对EC-1细胞增殖的抑制率递升(P<0.05或P<0.01)。随ARG浓度的递增,EC-1细胞凋亡率和凋亡指数递升(P均<0.01),Bcl-2表达率递降(P<0.01),Bax表达率递升(P<0.01)。上述变化与对照组比较,差异均有统计学意义(P<0.05或P<0.01)。结论 ARG可明显抑制EC-1细胞增殖并诱导其凋亡,其机制可能与ARG调控Bcl-2、Bax基因表达而诱导EC-1细胞凋亡有关。
Objective To observe the effect of arctigenin (ARG) on the proliferation and apoptosis of human esophageal cancer cells (EC-1) and to explore its mechanism. Methods ARG cells were treated with different concentrations of ARG. Cell proliferation was measured by MTT assay (ARG concentrations were 2.5, 5.0, 10.0, 20.0 and 40.0 mg / L, respectively. The duration of action was 24, 48 and 72 , 96,120h). The apoptosis rate was detected by flow cytometry. The apoptosis index was detected by agarose gel electrophoresis and DNA nick end labeling (TUNEL). The expressions of apoptosis-related genes Bcl-2 and Bax were detected by immunocytochemistry (The three kinds of test were used: ARG concentration 10.0,20.0,40.0mg / L, action time 24h); analysis of different concentrations of ARG on EC-1 cells. And set negative control group (only plus medium without drug) for comparison. Results The effect of ARG on EC-1 cell proliferation and apoptosis was dose-dependent. With the increase of ARG concentration and action time, the inhibition of EC-1 cell proliferation increased (P <0.05 or P <0.01). With the increasing of ARG concentration, the apoptosis rate and apoptosis index of EC-1 cells increased (P <0.01), the expression of Bcl-2 decreased (P <0.01) and the expression of Bax increased (P <0.01). The above changes compared with the control group, the difference was statistically significant (P <0.05 or P <0.01). Conclusion ARG can significantly inhibit the proliferation of EC-1 cells and induce its apoptosis. The mechanism may be related to ARG regulating the expression of Bcl-2 and Bax and inducing the apoptosis of EC-1 cells.